Neisseria gonorrhoeae is a strict human pathogen that causes an estimated ninety-five million cases of gonorrhea worldwide each year. Without an effective vaccine, antibiotic therapy remains the principle mechanism to reduce or block the transmission of the gonococcus in the community. Unfortunately, because of the development of mutations or the acquisition of resistance determinants, the gonococcus has developed resistance to relatively inexpensive antibiotics and new strategies will be needed to counteract this problem. Gonorrhea is an ancient disease of mankind. Thus, it is not surprising that this strict human pathogen has evolved mechanisms to deal with antimicrobial threats. We have identified a novel mechanism used by gonococci to resist the host-derived antimicrobials (e.g., mediators of innate host defense) that mimic the action of classical antibiotics; this resistance involves the coating of gonococci with host-derived polyamines. We have found that biologically relevant concentrations of polyamines can also inhibit biofilm formation by gonococci. Using spermine as a model polyamine, we have acquired evidence that gonococci mount a stress response in its presence. In order to better understand how polyamines influence the pathogenic potential of gonococci during infection, we will define bacterial factors that are important in determining polyamine-mediated modulation of biofilm formation and dispersion, as well as the interaction of gonococci with host cells (Specific Aim 1). We will also determine the responses made by gonococci at the transcriptional and translational levels in the presence of physiologically relevant levels of spermine and will construct nonpolar mutations in spermine-responsive genes so as to better understand how this pathogen might use host polyamines to proliferate during infection (Specific Aim 2). The results obtained during the two-year funding should provide novel insights regarding how the gonococcus responds to compounds in its natural environment, which is important in the development of new therapeutic agents that block infection or eradicate gonococci at the genital mucosal surface of men and women.